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Possible toxic effects of ofloxacin on human corneal endothelia were assessed by using electron microscopy and in vitro specular perfusion.Five pairs of corneas [with one cornea of each pair receiving balanced salt solution (Endosol; Allergan, Irvine. CA, U.S.A.) and the other receiving Endosol with 10 μg/ml of ofloxacin] underwent perfusion for 3 h with corneal pachymetry every 15 min followed by tissue fixation. A mean corneal swelling rate was calculated from a first-order regression line fit for each of the five experiments. The swelling rates between groups was compared by using a paired t test. Scanning and transmission electron micrographs were examined for cellular architecture. Another group of five pairs of corneas was perfused the same way with the only difference being a test dose of 30 μg/ml.The mean swelling rate for corneas perfused with 10 μg/ml of ofloxacin of −3.5 μm/h was not significantly different from that of the Endosol-alone corneas at −3.0 μm/h (p = 0.71). The mean swelling rate for corneas perfused at 30 μg/ml of ofloxacin was −4.1 μm/h, not significantly different from Endosol-alone perfused corneas at −6.5 μm/h (p = 0.08). No consistent ultra-structural changes could be attributed to exposure to 10 or 30 μg/ml of ofloxacin.Human corneal endothelium can be exposed to ofloxacin at a dose of 30 μg/ml for a period of 3 h without adverse ultrastructural or physiologic side effects.